Small molecule inhibition of transcription factor SALL4 and uses thereof

ABSTRACT

Provided herein are compounds that interrupt the function of SALL4. Also described are pharmaceutical compositions and medical uses of these compounds.

RELATED APPLICATION

This application claims the benefit of priority to U.S. ProvisionalPatent Application Ser. No. 62/567,939, filed Oct. 4, 2017, which ishereby incorporated herein by reference in its entirety.

BACKGROUND

The embryonic stem cell gene SALL4 encodes a zinc finger transcriptionfactor. Its expression is down-regulated during development, and absentin most adult tissues, but aberrantly re-expressed in cancer cells,including most AMLs. SALL4 is enriched in a “side-population (SP)” ofthe tumor cells. The SP is implicated in drug resistance and cancerinitiation, and is used to isolate the cancer initiation cells (CICs).SALL4 expression is correlated with worse prognosis in AML patients aswell. Knocking down the SALL4 gene by shRNA in leukemia leads to celldeath and growth inhibition both in vitro and in vivo. However,targeting transcription factor DNA binding ability and direct inhibitionof SALL4 remain challenging, as there are no reported SALL4 inhibitors.

Thus, there is a continuing need for pharmacologic agents that interruptthe function of SALL4 and that can be used to manipulate SALL4 intherapeutic or experimental applications.

SUMMARY OF INVENTION

In one aspect, the invention relates to compounds having the structureof Formula I or a pharmaceutically acceptable salt thereof:

wherein R¹, R², and Z are defined herein.

In another aspect, the invention relates to pharmaceutical compositionsof a compound of Formula I and a pharmaceutically acceptable carrier.

The invention also relates to methods of treating or preventing adisease or condition comprising administering a compound or compositionof the invention. In certain embodiments, the disease is cancer. Theinvention further relates to methods of inhibiting proliferation of acancer cell, comprising contacting a cancer cell with a compound orcomposition of the invention.

DETAILED DESCRIPTION OF THE INVENTION

In certain aspects, the invention provides various novel compounds, andpharmaceutical compositions thereof. In particular, such compounds areuseful as SALL4 inhibitors, and thus can be used to treat or prevent adisease or condition (e.g., cancer).

I. Compounds

In certain embodiments, the invention relates to compounds having thestructure of Formula I, or a pharmaceutically acceptable salt thereof:

wherein

Z is N or CR³;

R¹ is optionally substituted alkyl, cycloalkyl, heterocyclyl, aryl, orheteroaryl;

R² is optionally substituted alkyl, cycloalkyl, heterocyclyl, aryl, orheteroaryl; and

R³ is H or optionally substituted alkyl, carboxy or ester.

In certain embodiments, is N. In certain other embodiments, Z is CR³. Incertain embodiments, R³ is H. In certain other embodiments, R³ is ester(e.g.,

In certain embodiments, R¹ is optionally substituted aryl (e.g.,phenyl). In some embodiments, the aryl or phenyl is substituted with onemore groups selected from halo, hydroxyl, boronic acid, cyano,optionally substituted alkyl, optionally substituted alkoxy, carboxy,amino, ester, and optionally substituted aryl. For example, in certainembodiments, R¹ is selected from

In certain embodiments, R¹ is selected from optionally substitutednapthalenyl. In some embodiments, the napthalenyl is substituted withone or more groups selected from hydroxyl and nitro. For example, incertain embodiments, R¹ is selected from

In certain embodiments, R¹ is optionally substituted alkyl, cycloalkyl,or heterocyclyl.

In certain embodiments, the alkyl is substituted with one or more groupsselected from hydroxyl, alkoxy, optionally substituted alkyl, optionallysubstituted cycloalkyl, and optionally substituted aryl. For example, incertain embodiments, R¹ is selected from

In certain embodiments, cycloalkyl comprises 3 to 8 carbon atoms. Incertain embodiments, heterocyclyl comprises a nitrogen atom. Forexample, in certain embodiments, R¹ is selected from

In certain embodiments, the optionally substituted heteroaryl isselected from oxazolyl, pyrazolyl, imidazolyl, indolyl, benzoisoxazolyl,indazolyl, azaindolyl, benzothiazolyl, thiazolyl, thiophenyl, furanyl,pyridinyl, pyrmidinyl, pyrrolyl, quinolinyl, and carbazolyl.

In certain embodiments, the heteroaryl is substituted with one or moregroups selected from halo, oxy, nitro, sulfonate, and optionallysubstituted alkyl.

For example, in certain embodiments, R¹ is selected from

In certain embodiments, R² is optionally substituted alkyl, cycloalkyl,or aryl. In certain embodiments, alkyl is substituted with alkoxy, aminoor optionally substituted aryl. For example, in certain embodiments, R²is selected from

In certain embodiments, cycloalkyl comprises 3 to 6 carbon atoms. Forexample, in certain embodiments, R² is selected from

In certain embodiments, wherein R² is optionally substituted aryl (e.g.,phenyl). In certain embodiments, phenyl is substituted with one moregroups selected from halo, hydroxyl, cyano, optionally substitutedalkyl, optionally substituted alkoxy, carboxy, and ester. For example,in certain embodiments, R² is selected from

In certain embodiments, compounds of the invention may be racemic. Incertain embodiments, compounds of the invention may be enriched in oneenantiomer. For example, a compound of the invention may have greaterthan 30% ee, 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee, or even 95%or greater ee. The compounds of the invention have more than onestereocenter. Consequently, compounds of the invention may be enrichedin one or more diastereomer. For example, a compound of the inventionmay have greater than 30% de, 40% de, 50% de, 60% de, 70% de, 80% de,90% de, or even 95% or greater de.

In certain embodiments, as will be described in detail below, thepresent invention relates to methods of treating or preventing a diseaseor condition with a compound of Formula I, or a pharmaceuticallyacceptable salt thereof. In certain embodiments, the therapeuticpreparation may be enriched to provide predominantly one enantiomer of acompound of one of Formula I. An enantiomerically enriched mixture maycomprise, for example, at least 60 mol percent of one enantiomer, ormore preferably at least 75, 90, 95, or even 99 mol percent. In certainembodiments, the compound enriched in one enantiomer is substantiallyfree of the other enantiomer, wherein substantially free means that thesubstance in question makes up less than 10%, or less than 5%, or lessthan 4%, or less than 3%, or less than 2%, or less than 1% as comparedto the amount of the other enantiomer, e.g., in the composition orcompound mixture. For example, if a composition or compound mixturecontains 98 grams of a first enantiomer and 2 grams of a secondenantiomer, it would be said to contain 98 mol percent of the firstenantiomer and only 2% of the second enantiomer.

In certain embodiments, the therapeutic preparation may be enriched toprovide predominantly one diastereomer of a compound of one of FormulaI. A diastereomerically enriched mixture may comprise, for example, atleast 60 mol percent of one diastereomer, or more preferably at least75, 90, 95, or even 99 mol percent.

In certain embodiments, the present invention provides a pharmaceuticalpreparation suitable for use in a human patient in the treatment of adisease or condition, comprising an effective amount of any compound ofone of Formula I, and one or more pharmaceutically acceptableexcipients. In certain embodiments, the pharmaceutical preparations maybe for use in treating or preventing a condition or disease as describedherein. In certain embodiments, the pharmaceutical preparations have alow enough pyrogen activity to be suitable for use in a human patient.

Compounds of any of the above structures may be used in the manufactureof medicaments for the treatment of any diseases or conditions disclosedherein.

Exemplary compounds of the invention are depicted in Tables 1, 2, 3, and4. The compounds of Tables 1, 2, 3, and 4 are understood to encompassboth the free base and the conjugate acid. For example, the compounds inTables 1, 2, 3, and 4 may be depicted as complexes or salts withtrifluoroacetic acid or hydrochloric acid, but the compounds in theircorresponding free base forms or as salts with other acids are equallywithin the scope of the invention. Compounds may be isolated in eitherthe free base form, as a salt (e.g., a hydrochloride salt) or in bothforms. In the chemical structures shown below, standard chemicalabbreviations are sometimes used.

TABLE 1 Compounds of Library 1 Compound

B2

B3

C2

C3

D2

D3

E2

E3

F2

F3

G2

G3

B4

B5

C4

C5

D4

D5

E4

E5

F4

F5

G4

G5

B6

B7

C6

C7

D6

D7

E6

E7

F6

F7

G6

G7

B8

B9

C8

C9

D8

D9

E8

E9

F8

F9

G8

G9

B10

B11

C10

C11

D10

D11

E10

E11

F10

F11

G10

G11

TABLE 2 Exemplary of Library 2 Compound

B2

B3

C2

C3

D2

D3

E2

E3

F2

F3

G2

G3

B4

B5

C4

C5

D4

D5

E4

E5

F4

F5

G4

G5

B6

B7

C6

C7

D6

D7

E6

E7

F6

F7

G6

G7

B8

B9

C8

C9

D8

D9

E8

E9

F8

F9

G8

G9

B10

B11

C10

C11

D10

D11

E10

E11

F10

F11

G10

G11

TABLE 3 Compounds of Library 3 Compound DMSO A1

A2

B1

B2

C1

C2

D1

D2

E1

E2

F1

F2

G1

G2

H1

H2

A3

A4

B3

B4

C3

C4

D3

D4

E3

E4

F3

F4

G3

G4

H3

H4

A5

A6

B5

B6 DMSO C5

C6

D5

D6

E5

E6

F5

F6

G5

G6

H5

H6

A7

A8

B7

B8

C7

C8

D7

D8

E7

E8

F7

F8 DMSO G7

G8

H7

H8

A9

A10

B9

B10

C9

C10 Blank D9

D10

E9

E10

F9

F10

G9

G10

H9

H10

A11

A12

B11

B12

C11

C12

D11

D12

E11

E12 DMSO F11

F12

G11

G12

H11

H12

TABLE 4 Compounds of Library 4 Compound

A1

A2

B1

B2

C1

C2

D1

D2

E1

E2

F1

F2

G1

G2

H1

H2

A3

A4

B3

B4

C3

C4

D3

D4

E3

E4

F3

F4

G3

G4

H3

H4

A5

A6

B5

B6

C5

C6

D5

D6

E5

E6

F5

F6

G5

G6

H5

H6

A7

A8

B7

B8

C7

C8

D7

D8

E7

E8

F7

F8

G7

G8

H7

H8

A9

A10

B9

B10

C9

C10

D9

D10

E9

E10

F9

F10

G9

G10

H9

H10

A11

A12

B11

B12

C11

C12

D11

D12

E11

E12

F11

F12

G11

G12

H11

H12II. Pharmaceutical Compositions

In certain embodiments, the present invention provides pharmaceuticalcompositions comprising a compound of Formula I and a pharmaceuticallyacceptable carrier.

The compositions and methods of the present invention may be utilized totreat an individual in need thereof. In certain embodiments, theindividual is a mammal such as a human, or a non-human mammal. Whenadministered to an animal, such as a human, the composition or thecompound is preferably administered as a pharmaceutical compositioncomprising, for example, a compound of the invention and apharmaceutically acceptable carrier. Pharmaceutically acceptablecarriers are well known in the art and include, for example, aqueoussolutions such as water or physiologically buffered saline or othersolvents or vehicles such as glycols, glycerol, oils such as olive oil,or injectable organic esters. In a preferred embodiment, when suchpharmaceutical compositions are for human administration, particularlyfor invasive routes of administration (i.e., routes, such as injectionor implantation, that circumvent transport or diffusion through anepithelial barrier), the aqueous solution is pyrogen-free, orsubstantially pyrogen-free. The excipients can be chosen, for example,to effect delayed release of an agent or to selectively target one ormore cells, tissues or organs. The pharmaceutical composition can be indosage unit form such as tablet, capsule (including sprinkle capsule andgelatin capsule), granule, lyophile for reconstitution, powder,solution, syrup, suppository, injection or the like. The composition canalso be present in a transdermal delivery system, e.g., a skin patch.The composition can also be present in a solution suitable for topicaladministration, such as an eye drop.

A pharmaceutically acceptable carrier can contain physiologicallyacceptable agents that act, for example, to stabilize, increasesolubility or to increase the absorption of a compound such as acompound of the invention. Such physiologically acceptable agentsinclude, for example, carbohydrates, such as glucose, sucrose ordextrans, antioxidants, such as ascorbic acid or glutathione, chelatingagents, low molecular weight proteins or other stabilizers orexcipients. The choice of a pharmaceutically acceptable carrier,including a physiologically acceptable agent, depends, for example, onthe route of administration of the composition. The preparation orpharmaceutical composition can be a self-emulsifying drug deliverysystem or a self-microemulsifying drug delivery system. Thepharmaceutical composition (preparation) also can be a liposome or otherpolymer matrix, which can have incorporated therein, for example, acompound of the invention. Liposomes, for example, which comprisephospholipids or other lipids, are nontoxic, physiologically acceptableand metabolizable carriers that are relatively simple to make andadminister.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” as used herein means apharmaceutically acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial. Each carrier must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notinjurious to the patient. Some examples of materials which can serve aspharmaceutically acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalformulations.

A pharmaceutical composition (preparation) can be administered to asubject by any of a number of routes of administration including, forexample, orally (for example, drenches as in aqueous or non-aqueoussolutions or suspensions, tablets, capsules (including sprinkle capsulesand gelatin capsules), boluses, powders, granules, pastes forapplication to the tongue); absorption through the oral mucosa (e.g.,sublingually); anally, rectally or vaginally (for example, as a pessary,cream or foam); parenterally (including intramuscularly, intravenously,subcutaneously or intrathecally as, for example, a sterile solution orsuspension); nasally; intraperitoneally; subcutaneously; transdermally(for example as a patch applied to the skin); and topically (forexample, as a cream, ointment or spray applied to the skin, or as an eyedrop). The compound may also be formulated for inhalation. In certainembodiments, a compound may be simply dissolved or suspended in sterilewater. Details of appropriate routes of administration and compositionssuitable for same can be found in, for example, U.S. Pat. Nos.6,110,973, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, aswell as in patents cited therein.

The formulations may conveniently be presented in unit dosage form andmay be prepared by any methods well known in the art of pharmacy. Theamount of active ingredient which can be combined with a carriermaterial to produce a single dosage form will vary depending upon thehost being treated, the particular mode of administration. The amount ofactive ingredient that can be combined with a carrier material toproduce a single dosage form will generally be that amount of thecompound which produces a therapeutic effect. Generally, out of onehundred percent, this amount will range from about 1 percent to aboutninety-nine percent of active ingredient, preferably from about 5percent to about 70 percent, most preferably from about 10 percent toabout 30 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association an active compound, such as a compound ofthe invention, with the carrier and, optionally, one or more accessoryingredients. In general, the formulations are prepared by uniformly andintimately bringing into association a compound of the present inventionwith liquid carriers, or finely divided solid carriers, or both, andthen, if necessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules (including sprinkle capsules and gelatin capsules),cachets, pills, tablets, lozenges (using a flavored basis, usuallysucrose and acacia or tragacanth), lyophile, powders, granules, or as asolution or a suspension in an aqueous or non-aqueous liquid, or as anoil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup,or as pastilles (using an inert base, such as gelatin and glycerin, orsucrose and acacia) and/or as mouth washes and the like, each containinga predetermined amount of a compound of the present invention as anactive ingredient. Compositions or compounds may also be administered asa bolus, electuary or paste.

To prepare solid dosage forms for oral administration (capsules(including sprinkle capsules and gelatin capsules), tablets, pills,dragees, powders, granules and the like), the active ingredient is mixedwith one or more pharmaceutically acceptable carriers, such as sodiumcitrate or dicalcium phosphate, and/or any of the following: (1) fillersor extenders, such as starches, lactose, sucrose, glucose, mannitol,and/or silicic acid; (2) binders, such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose and/or acacia; (3) humectants, such as glycerol; (4)disintegrating agents, such as agar-agar, calcium carbonate, potato ortapioca starch, alginic acid, certain silicates, and sodium carbonate;(5) solution retarding agents, such as paraffin; (6) absorptionaccelerators, such as quaternary ammonium compounds; (7) wetting agents,such as, for example, cetyl alcohol and glycerol monostearate; (8)absorbents, such as kaolin and bentonite clay; (9) lubricants, such atalc, calcium stearate, magnesium stearate, solid polyethylene glycols,sodium lauryl sulfate, and mixtures thereof; (10) complexing agents,such as, modified and unmodified cyclodextrins; and (11) coloringagents. In the case of capsules (including sprinkle capsules and gelatincapsules), tablets and pills, the pharmaceutical compositions may alsocomprise buffering agents. Solid compositions of a similar type may alsobe employed as fillers in soft and hard-filled gelatin capsules usingsuch excipients as lactose or milk sugars, as well as high molecularweight polyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions, such as dragees, capsules (including sprinkle capsules andgelatin capsules), pills and granules, may optionally be scored orprepared with coatings and shells, such as enteric coatings and othercoatings well known in the pharmaceutical-formulating art. They may alsobe formulated so as to provide slow or controlled release of the activeingredient therein using, for example, hydroxypropylmethyl cellulose invarying proportions to provide the desired release profile, otherpolymer matrices, liposomes and/or microspheres. They may be sterilizedby, for example, filtration through a bacteria-retaining filter, or byincorporating sterilizing agents in the form of sterile solidcompositions that can be dissolved in sterile water, or some othersterile injectable medium immediately before use. These compositions mayalso optionally contain opacifying agents and may be of a compositionthat they release the active ingredient(s) only, or preferentially, in acertain portion of the gastrointestinal tract, optionally, in a delayedmanner. Examples of embedding compositions that can be used includepolymeric substances and waxes. The active ingredient can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-described excipients.

Liquid dosage forms useful for oral administration includepharmaceutically acceptable emulsions, lyophiles for reconstitution,microemulsions, solutions, suspensions, syrups and elixirs. In additionto the active ingredient, the liquid dosage forms may contain inertdiluents commonly used in the art, such as, for example, water or othersolvents, cyclodextrins and derivatives thereof, solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol,polyethylene glycols and fatty acid esters of sorbitan, and mixturesthereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations of the pharmaceutical compositions for rectal, vaginal, orurethral administration may be presented as a suppository, which may beprepared by mixing one or more active compounds with one or moresuitable nonirritating excipients or carriers comprising, for example,cocoa butter, polyethylene glycol, a suppository wax or a salicylate,and which is solid at room temperature, but liquid at body temperatureand, therefore, will melt in the rectum or vaginal cavity and releasethe active compound.

Formulations of the pharmaceutical compositions for administration tothe mouth may be presented as a mouthwash, or an oral spray, or an oralointment.

Alternatively or additionally, compositions can be formulated fordelivery via a catheter, stent, wire, or other intraluminal device.Delivery via such devices may be especially useful for delivery to thebladder, urethra, ureter, rectum, or intestine.

Formulations which are suitable for vaginal administration also includepessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining such carriers as are known in the art to be appropriate.

Dosage forms for the topical or transdermal administration includepowders, sprays, ointments, pastes, creams, lotions, gels, solutions,patches and inhalants. The active compound may be mixed under sterileconditions with a pharmaceutically acceptable carrier, and with anypreservatives, buffers, or propellants that may be required.

The ointments, pastes, creams and gels may contain, in addition to anactive compound, excipients, such as animal and vegetable fats, oils,waxes, paraffins, starch, tragacanth, cellulose derivatives,polyethylene glycols, silicones, bentonites, silicic acid, talc and zincoxide, or mixtures thereof.

Powders and sprays can contain, in addition to an active compound,excipients such as lactose, talc, silicic acid, aluminum hydroxide,calcium silicates and polyamide powder, or mixtures of these substances.Sprays can additionally contain customary propellants, such aschlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, suchas butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the active compound in theproper medium. Absorption enhancers can also be used to increase theflux of the compound across the skin. The rate of such flux can becontrolled by either providing a rate controlling membrane or dispersingthe compound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.Exemplary ophthalmic formulations are described in U.S. Publication Nos.2005/0080056, 2005/0059744, 2005/0031697 and 2005/004074 and U.S. Pat.No. 6,583,124, the contents of which are incorporated herein byreference. If desired, liquid ophthalmic formulations have propertiessimilar to that of lacrimal fluids, aqueous humor or vitreous humor orare compatible with such fluids. A preferred route of administration islocal administration (e.g., topical administration, such as eye drops,or administration via an implant).

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

Pharmaceutical compositions suitable for parenteral administrationcomprise one or more active compounds in combination with one or morepharmaceutically acceptable sterile isotonic aqueous or nonaqueoussolutions, dispersions, suspensions or emulsions, or sterile powderswhich may be reconstituted into sterile injectable solutions ordispersions just prior to use, which may contain antioxidants, buffers,bacteriostats, solutes which render the formulation isotonic with theblood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents that delay absorption such as aluminum monostearate andgelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsulated matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissue.

For use in the methods of this invention, active compounds can be givenper se or as a pharmaceutical composition containing, for example, 0.1to 99.5% (more preferably, 0.5 to 90%) of active ingredient incombination with a pharmaceutically acceptable carrier.

Methods of introduction may also be provided by rechargeable orbiodegradable devices. Various slow release polymeric devices have beendeveloped and tested in vivo in recent years for the controlled deliveryof drugs, including proteinacious biopharmaceuticals. A variety ofbiocompatible polymers (including hydrogels), including bothbiodegradable and non-degradable polymers, can be used to form animplant for the sustained release of a compound at a particular targetsite.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions may be varied so as to obtain an amount of the activeingredient that is effective to achieve the desired therapeutic responsefor a particular patient, composition, and mode of administration,without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound or combination ofcompounds employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound(s) being employed, the duration of the treatment,other drugs, compounds and/or materials used in combination with theparticular compound(s) employed, the age, sex, weight, condition,general health and prior medical history of the patient being treated,and like factors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the therapeutically effective amount of thepharmaceutical composition required. For example, the physician orveterinarian could start doses of the pharmaceutical composition orcompound at levels lower than that required in order to achieve thedesired therapeutic effect and gradually increase the dosage until thedesired effect is achieved. By “therapeutically effective amount” ismeant the concentration of a compound that is sufficient to elicit thedesired therapeutic effect. It is generally understood that theeffective amount of the compound will vary according to the weight, sex,age, and medical history of the subject. Other factors which influencethe effective amount may include, but are not limited to, the severityof the patient's condition, the disorder being treated, the stability ofthe compound, and, if desired, another type of therapeutic agent beingadministered with the compound of the invention. A larger total dose canbe delivered by multiple administrations of the agent. Methods todetermine efficacy and dosage are known to those skilled in the art(Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13ed., 1814-1882, herein incorporated by reference).

In general, a suitable daily dose of an active compound used in thecompositions and methods of the invention will be that amount of thecompound that is the lowest dose effective to produce a therapeuticeffect. Such an effective dose will generally depend upon the factorsdescribed above.

If desired, the effective daily dose of the active compound may beadministered as one, two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms. In certain embodiments of the presentinvention, the active compound may be administered two or three timesdaily. In preferred embodiments, the active compound will beadministered once daily.

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

This invention includes the use of pharmaceutically acceptable salts ofcompounds of the invention in the compositions and methods of thepresent invention. The term “pharmaceutically acceptable salt” as usedherein includes salts derived from inorganic or organic acids including,for example, hydrochloric, hydrobromic, sulfuric, nitric, perchloric,phosphoric, formic, acetic, lactic, maleic, fumaric, succinic, tartaric,glycolic, salicylic, citric, methanesulfonic, benzenesulfonic, benzoic,malonic, trifluoroacetic, trichloroacetic, naphthalene-2-sulfonic, andother acids. Pharmaceutically acceptable salt forms can include formswherein the ratio of molecules comprising the salt is not 1:1. Forexample, the salt may comprise more than one inorganic or organic acidmolecule per molecule of base, such as two hydrochloric acid moleculesper molecule of compound of Formula I or Formula II. As another example,the salt may comprise less than one inorganic or organic acid moleculeper molecule of base, such as two molecules of compound of Formula I orFormula II per molecule of tartaric acid.

In further embodiments, contemplated salts of the invention include, butare not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammoniumsalts. In certain embodiments, contemplated salts of the inventioninclude, but are not limited to, L-arginine, benenthamine, benzathine,betaine, calcium hydroxide, choline, deanol, diethanolamine,diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine,N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine,magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium,1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine,and zinc salts. In certain embodiments, contemplated salts of theinvention include, but are not limited to, Na, Ca, K, Mg, Zn or othermetal salts.

The pharmaceutically acceptable acid addition salts can also exist asvarious solvates, such as with water, methanol, ethanol,dimethylformamide, and the like. Mixtures of such solvates can also beprepared. The source of such solvate can be from the solvent ofcrystallization, inherent in the solvent of preparation orcrystallization, or adventitious to such solvent.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1)water-soluble antioxidants, such as ascorbic acid, cysteinehydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfiteand the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate,butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT),lecithin, propyl gallate, alpha-tocopherol, and the like; and (3)metal-chelating agents, such as citric acid, ethylenediamine tetraaceticacid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

III. Uses of Compounds and Compositions

In certain aspects, the invention provides methods of treating orpreventing a disease or condition, comprising administering to a subjecta compound of one of Formula I, e.g., in a therapeutically effectiveamount or a composition comprising a compound of Formula I.

In some embodiments, the disease is cancer. In some embodiments, thecancer is selected from acute myeloid leukemia, liver cancer, lungcancer, and myelodysplastic syndromes (MDS).

In certain embodiments, the cancer is a solid tumor. For example, thesubject is generally one who has been diagnosed as having a canceroustumor or one who has been previously treated for a cancerous tumor(e.g., where the tumor has been previously removed by surgery). Thecancerous tumor may be a primary tumor and/or a secondary (e.g.,metastatic) tumor.

In certain embodiments, the subject is a mammal, e.g., a human.

In certain embodiments, the invention provides methods of inhibitingproliferation of a cancerous cell comprising contacting a cancerous cellwith an effective amount of a compound of one of Formula I or acomposition comprising a compound of one of Formula I.

IV. Definitions

The term “acyl” is art-recognized and refers to a group represented bythe general formula hydrocarbylC(O)—, preferably alkylC(O)—.

The term “acylamino” is art-recognized and refers to an amino groupsubstituted with an acyl group and may be represented, for example, bythe formula hydrocarbylC(O)NH—.

The term “acyloxy” is art-recognized and refers to a group representedby the general formula hydrocarbylC(O)O—, preferably alkylC(O)O—.

The term “alkoxy” refers to an alkyl group, preferably a lower alkylgroup, having an oxygen attached thereto. Representative alkoxy groupsinclude methoxy, —OCF₃, ethoxy, propoxy, tert-butoxy and the like.

The term “cycloalkyloxy” refers to a cycloakyl group having an oxygenattached thereto.

The term “alkoxyalkyl” refers to an alkyl group substituted with analkoxy group and may be represented by the general formulaalkyl-O-alkyl.

The term “alkylaminoalkyl” refers to an alkyl group substituted with analkylamino group.

The term “alkenyl”, as used herein, refers to an aliphatic groupcontaining at least one double bond and is intended to include both“unsubstituted alkenyls” and “substituted alkenyls”, the latter of whichrefers to alkenyl moieties having substituents replacing a hydrogen onone or more carbons of the alkenyl group. Such substituents may occur onone or more carbons that are included or not included in one or moredouble bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed below, except where stability isprohibitive. For example, substitution of alkenyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated.

An “alkyl” group or “alkane” is a straight chained or branchednon-aromatic hydrocarbon which is completely saturated. Typically, astraight chained or branched alkyl group has from 1 to about 20 carbonatoms, preferably from 1 to about 10 unless otherwise defined. Examplesof straight chained and branched alkyl groups include methyl, ethyl,n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl,pentyl and octyl. A C₁-C₆ straight chained or branched alkyl group isalso referred to as a “lower alkyl” group.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification, examples, and claims is intended to include both“unsubstituted alkyls” and “substituted alkyls”, the latter of whichrefers to alkyl moieties having substituents replacing a hydrogen on oneor more carbons of the hydrocarbon backbone. Such substituents, if nototherwise specified, can include, for example, a halogen, a hydroxyl, acarbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl),a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate),an alkoxyl, a phosphoryl, a phosphate, a phosphonate, a phosphinate, anamino, an amido, an amidine, an imine, a cyano, a nitro, an azido, asulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, asulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic orheteroaromatic moiety. It will be understood by those skilled in the artthat the moieties substituted on the hydrocarbon chain can themselves besubstituted, if appropriate. For instance, the substituents of asubstituted alkyl may include substituted and unsubstituted forms ofamino, azido, imino, amido, phosphoryl (including phosphonate andphosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl andsulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls(including ketones, aldehydes, carboxylates, and esters), —CF₃, —CN andthe like. Exemplary substituted alkyls are described below. Cycloalkylscan be further substituted with alkyls, alkenyls, alkoxys, alkylthios,aminoalkyls, carbonyl-substituted alkyls, —CF₃, —CN, and the like.

The term “C_(x-y)” when used in conjunction with a chemical moiety, suchas acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to includegroups that contain from x to y carbons in the chain. For example, theterm “C_(x-y)alkyl” refers to substituted or unsubstituted saturatedhydrocarbon groups, including straight-chain alkyl and branched-chainalkyl groups that contain from x to y carbons in the chain, includinghaloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc.C₀ alkyl indicates a hydrogen where the group is in a terminal position,a bond if internal. The terms “C_(2-y)alkenyl” and “C_(2-y)alkynyl”refer to substituted or unsubstituted unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double or triple bond respectively.

The term “alkylamino”, as used herein, refers to an amino groupsubstituted with at least one alkyl group.

The term “alkylthio”, as used herein, refers to a thiol groupsubstituted with an alkyl group and may be represented by the generalformula alkylS-.

The term “alkynyl”, as used herein, refers to an aliphatic groupcontaining at least one triple bond and is intended to include both“unsubstituted alkynyls” and “substituted alkynyls”, the latter of whichrefers to alkynyl moieties having substituents replacing a hydrogen onone or more carbons of the alkynyl group. Such substituents may occur onone or more carbons that are included or not included in one or moretriple bonds. Moreover, such substituents include all those contemplatedfor alkyl groups, as discussed above, except where stability isprohibitive. For example, substitution of alkynyl groups by one or morealkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups iscontemplated.

The term “amide”, as used herein, refers to a group

wherein each R¹⁰ independently represent a hydrogen or hydrocarbylgroup, or two R¹⁰ are taken together with the N atom to which they areattached complete a heterocycle having from 4 to 8 atoms in the ringstructure.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines and salts thereof, e.g., a moietythat can be represented by

wherein each R¹⁰ independently represents a hydrogen or a hydrocarbylgroup, or two R¹⁰ are taken together with the N atom to which they areattached complete a heterocycle having from 4 to 8 atoms in the ringstructure.

The term “aminoalkyl”, as used herein, refers to an alkyl groupsubstituted with an amino group.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group.

The term “aryl” as used herein include substituted or unsubstitutedsingle-ring aromatic groups in which each atom of the ring is carbon.Preferably the ring is a 5- to 7-membered ring, more preferably a6-membered ring. The term “aryl” also includes polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings wherein at least one of the rings is aromatic,e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls,cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groupsinclude benzene, naphthalene, phenanthrene, phenol, aniline, and thelike.

The term “carbamate” is art-recognized and refers to a group

wherein R⁹ and R¹⁰ independently represent hydrogen or a hydrocarbylgroup, such as an alkyl group, or R⁹ and R¹⁰ taken together with theintervening atom(s) complete a heterocycle having from 4 to 8 atoms inthe ring structure.

The terms “carbocycle”, and “carbocyclic”, as used herein, refers to asaturated or unsaturated ring in which each atom of the ring is carbon.The term carbocycle includes both aromatic carbocycles and non-aromaticcarbocycles. Non-aromatic carbocycles include both cycloalkane rings, inwhich all carbon atoms are saturated, and cycloalkene rings, whichcontain at least one double bond. “Carbocycle” includes 5-7 memberedmonocyclic and 8-12 membered bicyclic rings. Each ring of a bicycliccarbocycle may be selected from saturated, unsaturated and aromaticrings. Carbocycle includes bicyclic molecules in which one, two or threeor more atoms are shared between the two rings. The term “fusedcarbocycle” refers to a bicyclic carbocycle in which each of the ringsshares two adjacent atoms with the other ring. Each ring of a fusedcarbocycle may be selected from saturated, unsaturated and aromaticrings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, maybe fused to a saturated or unsaturated ring, e.g., cyclohexane,cyclopentane, or cyclohexene. Any combination of saturated, unsaturatedand aromatic bicyclic rings, as valence permits, is included in thedefinition of carbocyclic. Exemplary “carbocycles” include cyclopentane,cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene,1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene andadamantane. Exemplary fused carbocycles include decalin, naphthalene,1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane,4,5,6,7-tetrahydro-1H-indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles”may be substituted at any one or more positions capable of bearing ahydrogen atom.

A “cycloalkyl” group is a cyclic hydrocarbon which is completelysaturated. “Cycloalkyl” includes monocyclic and bicyclic rings.Typically, a monocyclic cycloalkyl group has from 3 to about 10 carbonatoms, more typically 3 to 8 carbon atoms unless otherwise defined. Thesecond ring of a bicyclic cycloalkyl may be selected from saturated,unsaturated and aromatic rings. Cycloalkyl includes bicyclic moleculesin which one, two or three or more atoms are shared between the tworings. The term “fused cycloalkyl” refers to a bicyclic cycloalkyl inwhich each of the rings shares two adjacent atoms with the other ring.The second ring of a fused bicyclic cycloalkyl may be selected fromsaturated, unsaturated and aromatic rings. A “cycloalkenyl” group is acyclic hydrocarbon containing one or more double bonds.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl groupsubstituted with a carbocycle group.

The term “carbonate” is art-recognized and refers to a group —OCO₂—R¹⁰,wherein R¹⁰ represents a hydrocarbyl group.

The term “carboxy”, as used herein, refers to a group represented by theformula —CO₂H.

The term “ester”, as used herein, refers to a group —C(O)OR¹⁰ whereinR¹⁰ represents a hydrocarbyl group.

The term “ether”, as used herein, refers to a hydrocarbyl group linkedthrough an oxygen to another hydrocarbyl group. Accordingly, an ethersubstituent of a hydrocarbyl group may be hydrocarbyl-O—. Ethers may beeither symmetrical or unsymmetrical. Examples of ethers include, but arenot limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethersinclude “alkoxyalkyl” groups, which may be represented by the generalformula alkyl-O-alkyl.

The terms “halo” and “halogen” as used herein means halogen and includeschloro, fluoro, bromo, and iodo.

The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to analkyl group substituted with a hetaryl group.

The term “heteroalkyl”, as used herein, refers to a saturated orunsaturated chain of carbon atoms and at least one heteroatom, whereinno two heteroatoms are adjacent.

The term “heteroalkylamino”, as used herein, refers to an amino groupsubstituted with a heteralkyl group.

The terms “heteroaryl” and “hetaryl” include substituted orunsubstituted aromatic single ring structures, preferably 5- to7-membered rings, more preferably 5- to 6-membered rings, whose ringstructures include at least one heteroatom, preferably one to fourheteroatoms, more preferably one or two heteroatoms. The terms“heteroaryl” and “hetaryl” also include polycyclic ring systems havingtwo or more cyclic rings in which two or more carbons are common to twoadjoining rings wherein at least one of the rings is heteroaromatic,e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls,cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroarylgroups include, for example, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, pyrazole, pyridine, benzimidazole, quinoline,isoquinoline, quinoxaline, quinazoline, indole, isoindole, indazole,benzoxazole, pyrazine, pyridazine, purine, and pyrimidine, and the like.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, andsulfur.

The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer tosubstituted or unsubstituted non-aromatic ring structures, preferably 3-to 10-membered rings, more preferably 3- to 7-membered rings, whose ringstructures include at least one heteroatom, preferably one to fourheteroatoms, more preferably one or two heteroatoms. The terms“heterocyclyl” and “heterocyclic” also include polycyclic ring systemshaving two or more cyclic rings in which two or more carbons are commonto two adjoining rings wherein at least one of the rings isheterocyclic, e.g., the other cyclic rings can be cycloalkyls,cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls.Heterocyclyl groups include, for example, piperidine, piperazine,pyrrolidine, morpholine, lactones, lactams, and the like. Heterocyclylgroups can also be substituted by oxo groups. For example,“heterocyclyl” encompasses both pyrrolidine and pyrrolidinone.

The term “heterocycloalkyl”, as used herein, refers to an alkyl groupsubstituted with a heterocycle group.

The term “heterocycloalkylamino”, as used herein refers to an aminogroup substituted with a heterocycloalkyl group.

The term “hydrocarbyl”, as used herein, refers to a group that is bondedthrough a carbon atom that does not have a ═O or ═S substituent, andtypically has at least one carbon-hydrogen bond and a primarily carbonbackbone, but may optionally include heteroatoms. Thus, groups likemethyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to behydrocarbyl for the purposes of this application, but substituents suchas acetyl (which has a ═O substituent on the linking carbon) and ethoxy(which is linked through oxygen, not carbon) are not. Hydrocarbyl groupsinclude, but are not limited to aryl, heteroaryl, carbocycle,heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

The term “hydroxyalkyl”, as used herein, refers to an alkyl groupsubstituted with a hydroxy group.

The term “lower” when used in conjunction with a chemical moiety, suchas, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant toinclude groups where there are ten or fewer non-hydrogen atoms in thesubstituent, preferably six or fewer. A “lower alkyl”, for example,refers to an alkyl group that contains ten or fewer carbon atoms,preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl,alkenyl, alkynyl, or alkoxy substituents defined herein are respectivelylower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, orlower alkoxy, whether they appear alone or in combination with othersubstituents, such as in the recitations hydroxyalkyl and aralkyl (inwhich case, for example, the atoms within the aryl group are not countedwhen counting the carbon atoms in the alkyl substituent).

As used herein, the term “oxo” refers to a carbonyl group. When an oxosubstituent occurs on an otherwise saturated group, such as with anoxo-substituted cycloalkyl group (e.g., 3-oxo-cyclobutyl), thesubstituted group is still intended to be a saturated group. When agroup is referred to as being substituted by an “oxo” group, this canmean that a carbonyl moiety (i.e., —C(═O)—) replaces a methylene unit(i.e., —CH₂—).

The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two ormore rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls,heteroaryls, and/or heterocyclyls) in which two or more atoms are commonto two adjoining rings, e.g., the rings are “fused rings”. Each of therings of the polycycle can be substituted or unsubstituted. In certainembodiments, each ring of the polycycle contains from 3 to 10 atoms inthe ring, preferably from 5 to 7.

The term “silyl” refers to a silicon moiety with three hydrocarbylmoieties attached thereto.

The term “substituted” refers to moieties having substituents replacinga hydrogen on one or more carbons of the backbone. It will be understoodthat “substitution” or “substituted with” includes the implicit provisothat such substitution is in accordance with permitted valence of thesubstituted atom and the substituent, and that the substitution resultsin a stable compound, e.g., which does not spontaneously undergotransformation such as by rearrangement, cyclization, elimination, etc.As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and non-aromaticsubstituents of organic compounds. The permissible substituents can beone or more and the same or different for appropriate organic compounds.For purposes of this invention, the heteroatoms such as nitrogen mayhave hydrogen substituents and/or any permissible substituents oforganic compounds described herein which satisfy the valences of theheteroatoms. Substituents can include any substituents described herein,for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, analkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as athioester, a thioacetate, or a thioformate), an alkoxyl, a phosphoryl, aphosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine,an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, asulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, aheterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. Itwill be understood by those skilled in the art that substituents canthemselves be substituted, if appropriate. Unless specifically stated as“unsubstituted,” references to chemical moieties herein are understoodto include substituted variants. For example, reference to an “aryl”group or moiety implicitly includes both substituted and unsubstitutedvariants.

The term “sulfate” is art-recognized and refers to the group —OSO₃H, ora pharmaceutically acceptable salt thereof.

The term “sulfonamide” is art-recognized and refers to the grouprepresented by the general formulae

wherein R⁹ and R¹⁰ independently represents hydrogen or hydrocarbyl,such as alkyl, or R⁹ and R¹⁰ taken together with the intervening atom(s)complete a heterocycle having from 4 to 8 atoms in the ring structure.

The term “sulfoxide” is art-recognized and refers to the group—S(O)—R¹⁰, wherein R¹⁰ represents a hydrocarbyl.

The term “sulfonate” is art-recognized and refers to the group SO₃H, ora pharmaceutically acceptable salt thereof.

The term “sulfone” is art-recognized and refers to the group —S(O)₂—R¹⁰,wherein R¹⁰ represents a hydrocarbyl.

The term “thioalkyl”, as used herein, refers to an alkyl groupsubstituted with a thiol group.

The term “thioester”, as used herein, refers to a group —C(O)SR¹⁰ or—SC(O)R¹⁰ wherein R¹⁰ represents a hydrocarbyl.

The term “thioether”, as used herein, is equivalent to an ether, whereinthe oxygen is replaced with a sulfur.

The term “urea” is art-recognized and may be represented by the generalformula

wherein R⁹ and R¹⁰ independently represent hydrogen or a hydrocarbyl,such as alkyl, or either occurrence of R⁹ taken together with R¹⁰ andthe intervening atom(s) complete a heterocycle having from 4 to 8 atomsin the ring structure.

“Protecting group” refers to a group of atoms that, when attached to areactive functional group in a molecule, mask, reduce or prevent thereactivity of the functional group. Typically, a protecting group may beselectively removed as desired during the course of a synthesis.Examples of protecting groups can be found in Greene and Wuts,Protective Groups in Organic Chemistry, 3^(rd) Ed., 1999, John Wiley &Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods,Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogenprotecting groups include, but are not limited to, formyl, acetyl,trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert-butoxycarbonyl(“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl(“TES”), trityl and substituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (“FMOC”), nitro-veratryloxycarbonyl(“NVOC”) and the like. Representative hydroxyl protecting groupsinclude, but are not limited to, those where the hydroxyl group iseither acylated (esterified) or alkylated such as benzyl and tritylethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilylethers (e.g., TMS or TIPS groups), glycol ethers, such as ethyleneglycol and propylene glycol derivatives and allyl ethers.

As used herein, a therapeutic that “prevents” a disorder or conditionrefers to a compound that, in a statistical sample, reduces theoccurrence of the disorder or condition in the treated sample relativeto an untreated control sample, or delays the onset or reduces theseverity of one or more symptoms of the disorder or condition relativeto the untreated control sample.

The term “treating” includes prophylactic and/or therapeutic treatments.The term “prophylactic or therapeutic” treatment is art-recognized andincludes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic (i.e., it protects thehost against developing the unwanted condition), whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic, (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The term “prodrug” is intended to encompass compounds which, underphysiologic conditions, are converted into the therapeutically activeagents of the present invention (e.g., a compound of Formula I). Acommon method for making a prodrug is to include one or more selectedmoieties which are hydrolyzed under physiologic conditions to reveal thedesired molecule. In other embodiments, the prodrug is converted by anenzymatic activity of the host animal. For example, esters or carbonates(e.g., esters or carbonates of alcohols or carboxylic acids) arepreferred prodrugs of the present invention. In certain embodiments,some or all of the compounds of Formula I in a formulation representedabove can be replaced with the corresponding suitable prodrug, e.g.,wherein a hydroxyl in the parent compound is presented as an ester or acarbonate or carboxylic acid present in the parent compound is presentedas an ester.

EXAMPLES Example 1: Chemical Syntheses

The compounds disclosed herein were prepared via aGroebke-Blackburn-Bienayme Multicomponent Reaction as highlighted inScheme 1.

Library 1 Synthesis

Each amine (A1-A3), isocyanide (B1 and B2) and aldehyde (C1-C10) wasprepared as 400 mM DMSO solution. (See Table 5). Sc(OTf)₃ was preparedas a 20 mM DMSO solution. In each well on a 96 well plate, a differentcombination of 20 uL of the 400 mM solutions of an amine, isocyanide,aldehyde (1.0 equivalents each), as detailed by the plate map, andSc(OTf)₃ (0.05 equivalents) was combined to give final concentrations of100 mM product. To the outer wells of the plate were added 100 uL ofDMSO. Once all components were added to a plate, the plate was sealedand centrifuged at 1000×g for 30 s. The plate was then placed in an ovenat 110° C. overnight (15 h). Reactions were spot checked for productconversion by LCMS. Plate was submitted for assay testing.

TABLE 5 Reaction components for Library 1 Amines

Isocyanides

Aldehyde

Library 2 Synthesis

Each amine (A1-A3), isocyanide (B1 and B2) and aldehyde (C1-C10) wasprepared as 400 mM DMSO solution. Sc(OTf)₃ was prepared as a 20 mM DMSOsolution. (See Table 6). In each well on a 96 well plate, a differentcombination of 20 uL of the 400 mM solutions of an amine, isocyanide,aldehyde (1.0 equivalents each), as detailed by the plate map, andSc(OTf)₃ (0.05 equivalents) was combined to give final concentrations of100 mM product. To the outer wells of the plate were added 100 uL ofDMSO. Once all components were added to a plate, the plate was sealedand centrifuged at 1000×g for 30 s. The plate was then placed in an ovenat 110° C. overnight (15 h). Reactions were spot checked for productconversion by LCMS. Plate was submitted for assay testing.

TABLE 6 Reaction components for Library 2 Amines

Isocyanides

Aldehyde

Library 3 Synthesis

2-aminopyrazine (23.8 mg, 0.25 mmol) and Sc(OTf)₃ (6.2 mg, 0.013 mmol)were combined and suspended in DMSO (2.5 mL). To the reaction was thenadded cyclohexylisocyanide (31.1 uL, 0.25 mmol). The mixture was thenadded to a 96 well plate, adding 25 uL/well. To each well containing thereaction mixture, 25 uL of a 100 mM solution of an aldehyde was added.The plate was then sealed, centrifuged at 1000×g for 30 s and placed inan oven at 100° C. overnight (17 h). Reactions were spot checked forproduct conversion by LCMS. Plate was submitted for assay testing.

Library 4 Synthesis

Each amine (A1 and A2), isocyanide (I1-I12) and aldehyde (B1-B4) wasprepared as 400 mM DMSO solution. (See Table 7). Sc(OTf)₃ was preparedas a 20 mM DMSO solution. In each well on a 96 well plate, a differentcombination of 10 uL of the 400 mM solutions of an amine, isocyanide,aldehyde (1.0 equivalents each), as detailed by the plate map, andSc(OTf)₃ (0.05 equivalents) was combined to give final concentrations of100 mM product. Once all components were added to a plate, the plate wassealed and centrifuged at 1000×g for 30 s. The plate was then placed inan oven at 100° C. overnight. Reactions were spot checked for productconversion by LCMS. Plate was submitted for assay testing.

TABLE 7 Reaction components for Library 4 Amines

Isocyanides

Aldehyde

Library 1, Compound C10

3-methyl-1H-pyrazole-4-carbaldehyde (44.2 mg, 0.40 mmol) and2-aminopyridine (48.7 mg, 0.52 mmol) were combined and suspended in 3:1DCM/MeOH (2 mL). To the reaction was then added tert-butyl isocyanide(58.8 uL, 0.52 mmol) followed by Sc(OTf)₃ (10.0 mg, 0.02 mmol). Thereaction was heated via microwave to 110° C. for 30 minutes. Reactionwas filtered, washed with EtOAc and concentrated to dryness on therotovap. Crude compound was purified via ISCO chromatography (0-10%MeOH:DCM). Product yield: 16.8 mg, 15.6%

Library 1, Compound E10

3-methyl-1H-pyrazole-4-carbaldehyde (44.1 mg, 0.40 mmol) and2-aminopyrazine (49.6 mg, 0.52 mmol) were combined and suspended in 3:1DCM/MeOH (2 mL). To the reaction was then added tert-butyl isocyanide(58.8 uL, 0.52 mmol) followed by Sc(OTf)₃ (10.2 mg, 0.02 mmol). Thereaction was heated via microwave to 110° C. for 30 minutes. Reactionwas filtered, washed with EtOAc and concentrated to dryness on therotovap. Crude compound was purified via ISCO chromatography (0-10%MeOH:DCM). Product yield: 26.7 mg, 24.7%

Library 1, Compound D6

1H-indazole-5-carbaldehyde (58.7 mg, 0.40 mmol) and pyrazin-2-amine(38.2 mg, 0.40 mmol) were combined and suspended in DMSO (2 mL). To thereaction was then added isocyanocyclohexane (49.9 uL, 0.40 mmol)followed by HClO₄ (1.2 uL, 0.02 mmol). The reaction was heated to 100°C. for 2 hours. Reaction was quenched with H2O and desired productprecipitated out. Reaction was filtered and the crude precipitate waspurified via ISCO chromatography (0-100% EtOAc:Hexanes). Product yield:18.8 mg, 14.1%

Library 4. Compound A1

Pyridin-2-amine (0.40 mmol), 2-hydroxy-1-naphthaldehyde (0.80 mmol) andisocyanocyclopropane (0.15 mmol) were prepared as 400 mM solutions inDMSO and combined. Sc(OTf)₃ (0.003 mmol) was added as a 20 mM DMSOsolution to the reaction. Reaction was heated to 100° C. overnight.Reaction was purified via HPLC.

Library 4, Compound A12

Pyridin-2-amine (0.40 mmol), 2-hydroxy-1-naphthaldehyde (0.80 mmol) and2-isocyano-2-methylpropane (0.40 mmol) were prepared as 400 mM solutionsin DMSO and combined. Sc(OTf)₃ (0.003 mmol) was added as a 20 mM DMSOsolution to the reaction. Reaction was heated to 100° C. overnight.Reaction was purified via HPLC.

Library 4, Compound E1

Pyrazin-2-amine (0.40 mmol), 2-hydroxy-1-naphthaldehyde (0.80 mmol) andisocyanocyclopropane (0.15 mmol) were prepared as 400 mM solutions inDMSO and combined. Sc(OTf)₃ (0.003 mmol) was added as a 20 mM DMSOsolution to the reaction. Reaction was heated to 100° C. overnight.Reaction was purified via HPLC.

Library 4, Compound E2

Pyrazin-2-amine (0.40 mmol), 2-hydroxy-1-naphthaldehyde (0.80 mmol) and2-isocyano-3-methoxybenzene (0.44 mmol) were prepared as 400 mMsolutions in DMSO and combined. Sc(OTf)₃ (0.003 mmol) was added as a 20mM DMSO solution to the reaction. Reaction was heated to 100° C.overnight. Reaction was purified via HPLC.

Example 2

A biochemical assay was developed based on the AlphaScreen assay tomonitor the inhibition of SALL4. Table 8 highlights exemplary data fromthe AlphaScreen assay for compounds of Formula I. SRI 311 is a controlcompound.

TABLE 8 Assay Data from compounds of Library 4 Compound IC50 μM A1 0.1093 A4  2.231 A10 2.77 A11 1.506 A12 0.1616 E1  0.3347 E2  0.2876 E4 1.357 E5  0.6193 E6  0.4339 E7  2.489 E8  0.6226 E9  0.4976 E10 0.6157E11 0.5938 E12 0.7834 SRI 311 ~3.575

Example 3

The biological activity of lead compounds was assessed in a cellularassay. Table 9 highlights exemplary data from the WST8/Cell TiterViability Assay for compounds of Formula I.

TABLE 9 Assay Data for compounds of Library 4 WST8 WST8 Cell Titer CellTiter Compound H549 H611 H549 H611 A1  27.45 6.67 37.37 13.16 A4  38.1715.58 ~36.75 14.01 A10 >100 25.8 >100 27.39 A11 23.78 8.292 40.92 11.14A12 26.6 10.12 22.47 7.808 E1  >100 15.22 >100 19.4 E2  >100 17.82 >10019.56 E4  >100 23.53 >100 24.41 E5  >100 26.97 >100 41.43 E6  >10043.54 >100 ~42.85 E7  46.33 58.92 >100 ~40.18 E8  34.93 ~38.92 >100~41.47 E9  NA NA NA NA E10 18.66 20.95 >100 ~39.60 E11 75.14 98.13 >100~43.93 E12 >100 >100 >100 >100 A9  9.095 10.28 ~40.41 13.65 SRI 3113.203 3.448 29.26 8.444 SRI 027 25.19 29.66 30.08 29.71Vandy001 >100 >100 >100 >100

INCORPORATION BY REFERENCE

All publications and patents mentioned herein are hereby incorporated byreference in their entirety as if each individual publication or patentwas specifically and individually indicated to be incorporated byreference. In case of conflict, the present application, including anydefinitions herein, will control.

EQUIVALENTS

While specific embodiments of the subject invention have been discussed,the above specification is illustrative and not restrictive. Manyvariations of the invention will become apparent to those skilled in theart upon review of this specification and the claims below. The fullscope of the invention should be determined by reference to the claims,along with their full scope of equivalents, and the specification, alongwith such variations.

The invention claimed is:
 1. A compound having a structure of Formula Ior a pharmaceutically acceptable salt thereof:

wherein Z is CR³; R¹ is

optionally substituted cycloalkyl, heterocyclyl, aryl, oxazolyl,indolyl, benzoisoxazolyl, indazolyl, azaindolyl, benzothiazolyl,imidazo[1,2-a]pyridine, or thiazolyl, wherein the optionally substitutedaryl is optionally substituted phenanthrenyl or naphthalenyl substitutedwith one or more groups selected from hydroxyl and nitro; R² isoptionally substituted alkyl, cycloalkyl, heterocyclyl, aryl, orheteroaryl; and R³ is H or optionally substituted alkyl, carboxy orester, provided when R¹ is cycloalkyl, aryl, heterocyclyl, or heteroaryland R² is optionally substituted alkyl, R³ is not alkyl.
 2. The compoundof claim 1, wherein R³ is


3. The compound of claim 1, wherein R¹ is naphthalenyl substituted withone or more groups selected from hydroxyl and nitro.
 4. The compound ofclaim 3, wherein R¹ is selected from


5. The compound of claim 1, wherein R¹ is selected from


6. The compound of claim 1, wherein the oxazolyl, indolyl,benzoisoxazolyl, indazolyl, azaindolyl, benzothiazolyl,imidazo[1,2-a]pyridine, and thiazolyl is substituted with one or moregroups selected from halo, oxy, nitro, sulfonate, and optionallysubstituted alkyl.
 7. The compound of claim 1, wherein R¹ is selectedfrom


8. The compound of claim 1, wherein R² is alkyl substituted with alkoxy,amino or optionally substituted aryl.
 9. The compound of claim 1,wherein R² is selected from


10. The compound of claim 1, wherein R² is selected from


11. The compound of claim 1, wherein R² is optionally substitutedphenyl.
 12. The compound of claim 11, wherein the phenyl is substitutedwith one more groups selected from halo, hydroxyl, cyano, optionallysubstituted alkyl, optionally substituted alkoxy, carboxy, and ester.13. The compound of claim 1, wherein R² is selected from


14. A compound selected from

or a pharmaceutically acceptable salt thereof.
 15. A pharmaceuticalcomposition comprising the compound or a pharmaceutically acceptablesalt thereof of claim 1 and a pharmaceutically acceptable carrier.
 16. Amethod of treating acute myeloid leukemia, liver cancer, lung cancer, ormyelodysplastic syndrome (MDS) comprising administering to a subject thecompound or a pharmaceutically acceptable salt thereof of claim
 1. 17.The compound of claim 14, which is

or a pharmaceutically acceptable salt thereof.
 18. A pharmaceuticalcomposition comprising the compound or a pharmaceutically acceptablesalt thereof of claim 17 and a pharmaceutically acceptable carrier. 19.The method of claim 16, wherein the cancer is acute myeloid leukemia.20. The method of claim 16, wherein the cancer is liver cancer.
 21. Themethod of claim 16, wherein the cancer is lung cancer.
 22. The method ofclaim 16, wherein the cancer is myelodysplastic syndrome.